Format

Send to

Choose Destination
Methods Mol Biol. 2015;1346:47-54. doi: 10.1007/978-1-4939-2987-0_4.

Photocleavable DNA Barcoding Antibodies for Multiplexed Protein Analysis in Single Cells.

Author information

1
Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, CPZN 5206, Boston, MA, 02114, USA.
2
Center for Systems Biology, Massachusetts General Hospital, 185 Cambridge Street, CPZN 5206, Boston, MA, 02114, USA. rweissleder@mgh.harvard.edu.

Abstract

We describe a DNA-barcoded antibody sensing technique for single cell protein analysis in which the barcodes are photocleaved and digitally detected without amplification steps (Ullal et al., Sci Transl Med 6:219, 2014). After photocleaving the unique ~70 mer DNA barcodes we use a fluorescent hybridization technology for detection, similar to what is commonly done for nucleic acid readouts. This protocol offers a simple method for multiplexed protein detection using 100+ antibodies and can be performed on clinical samples as well as single cells.

KEYWORDS:

Antibody conjugation; DNA barcoding; Fine needle aspirates; Multiplexing; Pathway analysis; Proteomics; Single cell analysis

PMID:
26542714
DOI:
10.1007/978-1-4939-2987-0_4
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Springer
Loading ...
Support Center